Lab Notebook

20th July

Sriram & Calin

No colonies on the plates from Friday. Suspect either that the 300 rpm mixing during the heat shock might have killed the cells, or that we added the cells too soon after adding the antibiotic, before it had time to diffuse. Redid the transformations correcting these steps once again with the following parts:
For Conjugation:
BBa_I714031 - OriT-R
BBa_J23100 - strong promoter
BBa_E0840 - GFP generator
BBa_I13522 - pTet GFP
For Delay:
BBa_E0040 - GFP
BBa_C0040 - TetR
BBa_C0051 - cl
BBa_I14018 - pBla
BBa_I12006 - pLambclin
BBa_E1010 - mRFP1

Heat shock was done in the waterbath at 42°C for 1 min. Antibiotic was added while the agar was still liquid, mixed, then poured onto the plates to solidify.

A beaker of eppendorf have been sent to kitchen for autoclaving.

Weenink

I did the assembly of my first biobrick today! Protocol was according to the ginkgo bioworks assembly kit. The following biobricks were used:
Upstream: B0032
Downstream: K145015
Backbone: pSB1AK3
Transformation was done on the PCR machine (as a test)

Also, I inoculated 5 ml liquid culture (lb, 37degrees shaking incubator) of top10 cells for making them chemically competent.